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White House HIV/AIDS office again opens the floor -- and phones -- for community inputby Yusef NajafiPublished on May 20 2010

The White House Office of National AIDS Policy (ONAP) presented a tentative outline of a National HIV/AIDS Strategy during a community meeting in D.C. and a national conference call on Friday, May 14.

''What we're telling you is where we're at, but I think we can all recognize that this will evolve,'' said ONAP Director Jeffrey Crowley, addressing the crowd of about 60 people in the South Court Auditorium of the Eisenhower Executive Office Building.

The 72-page booklet distributed at the meeting, ''Community Ideas for Improving the Response to the Domestic HIV Epidemic,'' provided the outline that was crafted after ONAP conducted an Internet survey on its website, as well as hosted 14 community discussions around the country, including one at the University of D.C. on Sept. 21, 2009.

The work-in-progress strategy is broken down into three components: prevent new HIV infections, increase access to care and optimize health outcomes, and reduce HIV-related health disparities.

''We can evolve based on what we hear today, but also as we continue engaging with the [Presidential Advisory Council on HIV/AIDS (PACHA)] and also our federal partners,'' Crowley added, later opening up the floor to comments from the audience.

''What we hear today is not set in stone, but we want to give you an update that will really signal the direction that we're moving in.''

Jen Heitel Yakush, director for public policy at the Sexuality Information and Education Council of the United States (SIECUS), based in D.C., attended the meeting and said comprehensive sex education is key.

''We need to be talking about HIV prevention and STI prevention in addition to unintended pregnancy prevention, and the inter-related health needs of particularly LGBT young people,'' Yakush, who is bisexual, told Metro Weekly immediately following the hearing.

''I have a lot of hope for the National HIV/AIDS Strategy,'' she added. ''We know that it's going to be broad strokes. I hope there's going to be some detailed goals and measurements within the National HIV/AIDS Strategy. We also know that even in these tight fiscal times we need increased funding if we're really going to have an impact on the number of new HIV infections every year.''

Among other local attendees was Don Blanchon, CEO of the Whitman-Walker Clinic, though he did not speak at the event.

For more information about the Office of National AIDS Policy, visit whitehouse.gov/administration/eop/onap/.

Scripps research scientists determine structure of immune molecule that counteracts HIV strainsThe findings advance the effort to develop an AIDS vaccineLA JOLLA, CA ? June 1, 2010 ?In findings that contribute to efforts to design an AIDS vaccine, a team led by Scripps Research Institute scientists has determined the structure of an immune system antibody molecule that effectively acts against most strains of human immunodeficiency virus (HIV), the virus that causes AIDS.

The study, which is being published in an advance, online issue of the journal Proceedings of the National Academy of Sciences (PNAS) during the week of June 1, 2010, illuminates an unusual human antibody called PG16.

"This study advances the overall goal of how to design an HIV vaccine," said Scripps Research Professor Ian Wilson, who led the team with Dennis Burton, Scripps Research professor and scientific director of the International AIDS Vaccine Initiative (IAVI) Neutralizing Antibody Center at Scripps Research. "This antibody is highly effective in neutralizing HIV-1 and has evolved novel features to combat the virus."

The Problem with HIV

According to the World Health Organization's latest statistics, around 33 million people are living with HIV worldwide. During 2008 alone, more than 2 million men, women, and children succumbed to the disease and an estimated 2.7 million were infected with HIV. One of the most compelling medical challenges today is to develop a vaccine that will provide complete protection to someone who is later exposed to this virus.

HIV causes AIDS by binding to, entering, and ultimately leading to the death of T helper cells, which are immune cells that are necessary to fight off infections by common bacteria and other pathogens. As HIV depletes the body of T helper cells, common pathogens can become potentially lethal.

An effective HIV vaccine would induce antibodies (specialized immune system molecules) against the virus prior to exposure to the virus. Also called immunoglobulins, these antibodies would circulate through the blood, and track down and kill the virus.

Most of the antibodies that the body produces to fight HIV, however, are ineffective. The surface of the virus is cloaked with sugar molecules that prevent antibodies from slipping in and blocking the proteins the virus uses to latch onto a cell and infect it. To make matters more complicated, HIV is constantly mutating, so there are multiple HIV strains that antibodies elicited in any vaccine must be able to sense and destroy.

Nonetheless, while rare, broadly neutralizing antibodies against HIV do exist.

Last year, a team of scientists from IAVI, Scripps Research, Theraclone Sciences, and Monogram Biosciences published research from a systematic search for such antibodies among 2,000 volunteers. The study revealed two powerful new broadly neutralizing antibodies against HIV?PG9 and PG16, isolated from a volunteer in Africa.

"Hammerhead" Structure

Once the broadly neutralizing antibodies were discovered, the next challenge was to figure out how they worked. To shed light on this question, in the current study members of the Wilson lab turned to x-ray crystallography, a technique that can solve structures to exquisitely high resolution.

In x-ray crystallography, scientists manipulate a protein or some other molecule so that a crystal forms. This crystal is then placed in front of a beam of x-rays, which diffract when they strike the atoms in the crystal. Based on the pattern of diffraction, scientists can reconstruct the shape of the original molecule. The scientists succeeded in forming crystals of the active part of the PG16 antibody, and in reconstructing the structure from the data?with some surprising results.

"The antibody has a novel and really interesting subdomain that hasn't been seen before," said Research Associate Rob Pejchal, who is first author of the paper. "This subdomain, which we found plays a major role in the recognition and neutralization of HIV, has a different kind of antibody architecture. We like to call it the 'hammerhead' because it resembles the head of a hammerhead shark. It reaches out from the main part of the antibody and it has two flat ends on top."

Co-author Laura Walker, a graduate student in the Scripps Research Kellogg School of Science and Technology, added, "This hypervariable loop (CDR3) that forms the novel subdomain is also unusually long for an antibody. Almost all of the antibodies we know to be broadly neutralizing against HIV have one unusual feature or another."

Pejchal notes that the study also revealed that PG16 was sulfated, suggesting possible mechanisms of action not usually seen in antibodies this effective against HIV.

While the scientists were unsuccessful so far in crystallizing PG16's sister molecule PG9, they were able to glean insight into its action from biochemical studies using both molecules. By switching a small (seven-amino acid) segment of the CDR3 subdomain of PG9 for a similar segment from PG16, the team changed the subset of HIV isolates neutralized by the antibody. This confirmed the loop in question was the "business end" of the antibody and suggested that it might be possible to create other interesting variants of the antibody by manipulating this region.

Seth Berkley, president and CEO of IAVI, which funded the study with the National Institute of Allergy and Infectious Diseases (NIAID) of the National Institute of Health (NIH), noted, "These studies of PG16 have taught us a lot about how these neutralizing antibodies work. I am particularly excited by the possibilities these findings open up for AIDS vaccine development, since the breadth and potency of HIV neutralization achieved by PG16 is what we'd like to see in the antibodies elicited by a vaccine. IAVI and its researchers will continue to support the application of these findings to the design of novel immunogens against HIV. We hope that we will be able to translate the insights gleaned from this study into the design of a promising AIDS vaccine candidate."

###

In addition to Wilson, Burton, Pejchal, and Walker, authors of the paper, "Crystal structure and functional studies of broadly reactive antibody PG16 reveal a novel H3 subdomain that mediates potent neutralization of HIV-1," are Robyn Stanfield and Pascal Poignard of Scripps Research and IAVI, Wayne Koff and Sanjay Phogat of IAVI New York. This study was supported by IAVI, NIAID, the Skaggs Institute of the Scripps Research Institute, and the Ragon Institute.

About The Scripps Research Institute

The Scripps Research Institute is one of the world's largest independent, non-profit biomedical research organizations, at the forefront of basic biomedical science that seeks to comprehend the most fundamental processes of life. Scripps Research is internationally recognized for its discoveries in immunology, molecular and cellular biology, chemistry, neurosciences, autoimmune, cardiovascular, and infectious diseases, and synthetic vaccine development. Established in its current configuration in 1961, it employs approximately 3,000 scientists, postdoctoral fellows, scientific and other technicians, doctoral degree graduate students, and administrative and technical support personnel. Scripps Research is headquartered in La Jolla, California. It also includes Scripps Florida, whose researchers focus on basic biomedical science, drug discovery, and technology development. Scripps Florida is located in Jupiter, Florida.

Getting Pregnant With HIV+ Husband. Barrier to Sperm Washing in California, A perilous road to reproduction

The Calif governor’s surprising veto of a fertility-procedure bill means HIV-discordant couples must risk their own health in order to conceive.By Katie Worth, Illustration by Matthew Hollister

July 1 2010, http://www.sanfranmag.comThis month, for the first time in her eight-year marriage, Deanna Crandell plans to have unprotected sex with her husband. It’s not a decision she made lightly: Her husband is HIV-positive and she is not, so the consequences could be grave. But Crandell, 34, is desperate to have a baby, and her only other option is a fertility procedure that most clinics aren’t willing to perform, despite the fact that it’s been legal in California for more than two years.

The method, called “sperm washing,” separates potentially infected semen from healthy sperm cells, making fertility treatments like in-vitro fertilization and intrauterine insemination safe for HIV-discordant couples like the Crandells. Until 2007, it was illegal in California because of an 18-year-old law that prohibited transferring tissue from an HIV-infected person to a healthy individual. But studies in the United States and Europe had recorded more than 4,500 successful sperm-washing inseminations without a single case of HIV transmission—so San Francisco state senator Carole Migden drafted a bill that would allow clinics to perform the procedure. It breezed through the state legislature and was signed by Governor Schwarzenegger.

Before advocates could celebrate, though, it became clear that the law contained at least one fatal flaw: It required the clinic that performed the procedure to provide post-treatment care, like administering antiretroviral drugs to further reduce a woman’s chances of contracting HIV. Most fertility specialists haven’t been trained in HIV care, so they’ve been reluctant to take on this responsibility—in fact, only one clinic in California offers the procedure.

In an effort to mend the ineffectual law, state assemblymember Jerry Hill, of San Mateo County, introduced a bill that essentially relieved clinics of the burden to provide post-treatment care. It was unanimously approved by the legislature and sent to the governor’s desk in September 2009. But something unexpected happened: Schwarzenegger vetoed it. He said he supported “the intent” of the bill, but he thought it would inappropriately restrict the California Department of Public Health. Senator Hill was struck dumb, especially since he had vetted the bill with the governor’s office and the CDPH, and neither had raised any red flags. “Families are running out of time here,” Hill says.

The Crandells, for instance, are no longer willing to wait. While they’ve saved enough money to be able to pay for the fertility procedure if it were offered locally, the couple cannot sustain repeated trips either out of state or to Los Angeles (where California’s only clinic willing to carry out HIV sperm washing is located) for the insemination plus the requisite follow-up appointments. So in October, Crandell met with doctors to prepare for her first attempt at timed unprotected sex. She knows she faces the possibility of contracting HIV (by one analysis, a 4.3 percent chance), but she feels it’s worth the risk. “We’ve been married for eight years,” she says. “We were ready to have kids eight years ago, and we’re more than ready now.”

Powered by TranslateContributed by Alton Parrish (Editor) Saturday, June 12, 2010 1:24 More stories from this contributorThis story has been viewed 80 times(27 times in the past 24 hours, 5 times in the past hour)1 person on this page right nowResearchers at the University of Oklahoma Health Sciences Center have found a way, for the first time, to create a vaccine using a protein that activates a distinct part of the immune system. The research has potential treatment and prevention applications for several viral diseases, cancer, tuberculosis and HIV.

The work appears in a recent issue of The Journal of Immunology.

"No one has ever done this with a T-cell vaccine, so we’re learning; but now we are starting to get some traction. We are finding that a T-cell vaccine can work," said William Hildebrand, Ph.D., the lead researcher on the project.

For years, Hildebrand and his research team have been working with the body’s alarm system to learn how cells alert the immune system that something is wrong. The goal is to create viable targets for vaccines that activate T-cells in the immune system. T-cells are responsible for killing virus-infected cells in your body. T-cells also kill cells that become cancerous. Some vaccines such as the smallpox vaccine activate T-cells, but this occurs inadvertently. Until now, vaccines have focused on generating antibodies to keep you from getting sick.

While many of these antibody (B-cell) vaccines work well, the dependence on antibodies has prompted some viruses to skirt antibody immunity, making vaccines less effective or not effective at all for some viruses. With a T-cell vaccine, researchers would be able to activate another arm of the immune system to target a specific virus in the body and kill it.

To develop the vaccine, Hildebrand began by determining how the immune system distinguishes between a virus-infected or cancerous cell and a healthy cell. Researchers determine what unique proteins are sent to the surface of the infected cell to provide T-cells with an optimal target. Different viruses send different proteins.

Researchers started with West Nile virus since West Nile doesn’t change like the flu or develop resistance like cancer or HIV. After developing the target over several years, researchers at the OU Health Sciences Center worked with colleagues at Washington University in St. Louis to create a vaccine.

The process is now being repeated for targets and vaccines in other areas, such as cancer, where activating T-cells is difficult.

"Now that we have demonstrated the feasibility of developing a T-cell-specific vaccine, we intend to use the same process to discover other reliable targets, validate them and develop additional vaccines," Hildebrand said.

The research is funded by grants and contracts totaling $15 million from the National Institutes of Health.

Australian researchers pave way for new vaccine15 June 2010 | by Nick O'Donoghue .Results of research carried out by the University of Sydney could help develop a vaccine to prevent Herpes Simplex Virus (HSV).

Findings published in the Journal of Immunology identified how skin cells are infected by the virus, which is associated with the promotion and transmission of Human Immunodeficiency Virus (HIV).

Lead investigator Associated Professor Cheryl Jones said HSV was a medically significant virus that caused devastating disease throughout life for sufferers.

“Potentially, we may be on the right track for a new discovery.

“The skin represents a major entry point; therefore understanding how immune cells behave during the infection is of vital importance to researchers trying to find a cure for HSV.

“HSV infection of the skin and genital mucosa are important for the promotion and transmission of HIV, the virus that causes AIDS,” she said.

The research carried out in mice found HSV can infect and kill Langerhans cells, the immune cell in the top layer of the skin, which should exit the skin to the lymph nodes to trigger a stronger immune response to infection.

Perinatal transmission of HIV, including during breast-feeding, can be markedly reduced by antiretroviral therapy, either of mothers or their infants.

That's the message from two randomized trials in Africa that compared various ways of using antiretroviral medications to prevent perinatal transmission, both reported in the June 17 issue of the New England Journal of Medicine.

Despite differences between the trials, taken together they imply that "it should be possible to eliminate new perinatal HIV-1 infections globally" with the appropriate use of the medications, said Lynne Mofenson, MD, of the NIH.

Action Points

* Explain to interested patients that these studies suggest that a range of interventions can reduce the risk that HIV-positive mothers will pass the virus to their newborns.

In an accompanying editorial, Mofenson argued that debate over which regimen is best "should not be used to justify inaction."

Instead, Mofenson said, the choice of regimen is secondary to identifying, caring for, and treating HIV-positive women and their infants.

The two trials used different medications, took place in different countries, and had differing goals.

A study in Malawi, led by Charles van der Horst, MD, of the University of North Carolina in Chapel Hill, looked at the breast-feeding period and asked whether treating the mother or the infant would have better results.

In Botswana, a team led by Roger Shapiro, MD, of Beth Israel Deaconess Medical Center in Boston, compared the effects on transmission of two regimens of maternal triple-drug therapy starting in the third trimester and extending during a six-month breast-feeding period.

In the Malawi study, all mothers and children got the now-standard perinatal prophylaxis of a single dose of nevirapine (Viramune) and a week of nevirapine and lamivudine (3TC). The 2,369 HIV-positive breast-feeding mothers all had a CD4-positive lymphocyte count of at least 250 cells per cubic millimeter and thus -- by then-current standards -- did not require antiretroviral therapy.

After the perinatal intervention, they were randomly assigned to no further treatment (the control group), to maternal therapy with a triple-drug regimen, or to infant treatment with nevirapine. The intervention period lasted 28 weeks.

The researchers found:

* 5% of infants were HIV-positive at two weeks.

* The estimated risk of HIV transmission between then and the end of the study period was 5.7% in the control group, compared with 2.9% in the maternal-regimen group (difference significant at P=0.009), and 1.7% in the infant-regimen group (significant at P<0.001).

* The estimated risk of the combined endpoint of infant HIV infection or death in the same period was 7.0% in the control group, 4.1% in the maternal-regimen group (difference significant at P=0.02), and 2.6% in the infant-regimen group (significant at P<0.001).

* 6.2% of women getting antiretroviral therapy suffered from neutropenia, compared with 2.6% of those whose infants were treated and 2.3% in the control group.

* 1.9% of the infants receiving nevirapine had a hypersensitivity reaction to the drug.

In the Botswana study, 560 pregnant HIV-positive women with a CD4 cell count of at least 200 per cubic millimeter were randomly assigned to get one of two triple-drug regimens, starting between 26 and 34 weeks gestation.

One group (dubbed the nucleoside reverse-transcriptase inhibitor [NRTI] group) got coformulated abacavir (Ziagen), zidovudine (AZT), and lamivudine, while the other (the protease-inhibitor group) got lopinavir-ritonavir (Kaletra) as well as zidovudine and lamivudine.

The control group consisted of 170 women with CD4 counts less than 200 cells per cubic millimeter who were treated with nevirapine, zidovudine, and lamivudine.

All infants got a single dose of nevirapine perinatally, followed by four weeks of zidovudine.

The researchers found:

* Viral control -- less than 400 copies of HIV RNA per milliliter -- did not differ significantly among the groups at delivery: 96% in the NRTI group, 93% in the protease-inhibitor group, and 94% in the control group.

* Viral control remained high and did not differ throughout the breast-feeding period: 92% in the NRTI group, 93% in the protease-inhibitor group, and 95% in the control group.

* By six months, eight of the 709 live-born infants (1.1%) were infected. Six were infected in utero (four in the NRTI group, and one each in the other groups), and two were infected during breast-feeding (both in the NRTI group).

* Side effects that limited treatment were seen in 2% of women in the NRTI group, 2% of women in the protease-inhibitor group, and 11% of women in the control group.

Although the trials can't be compared directly because of their different designs, Mofenson said some lessons can be learned.

One such lesson, she said, is that antiretroviral regimens aimed at preventing transmission should start during pregnancy. In the Malawi study, which did not treat women during pregnancy, in utero transmission was 5%, compared with 0.9% in the Botswana study.

The Malawi study also showed that highest risk for transmission was early in the breast-feeding period; during that period, the infant regimen reduced transmission by 86.1% (compared with the control group) and the maternal regimen reduced transmission by 52.8%.

The difference, Mofenson said, is probably a factor of the time the maternal regimen took to have an effect, suggesting that infant treatment is likely to be more important if the mother presents late in her pregnancy or during labor.

The Malawi study was supported by the Centers for Disease Control and Prevention, the National Institute of Allergy and Infectious Diseases, the University of North Carolina Center for AIDS Research, the NIH Fogarty AIDS International Training and Research Program, Abbott Laboratories, GlaxoSmithKline, Boehringer Ingelheim, Roche Pharmaceuticals, Bristol-Myers Squibb, the Elizabeth Glaser Pediatric AIDS Foundation, the United Nations Children's Fund, the World Food Program, the Malawi Ministry of Health and Population, Johnson & Johnson, and the U.S. Agency for International Development.

* Explain to interested patients that this study suggests that drinking diet colas may be associated with a negative calcium balance, leaving people at risk for fracture.

* Note that this was a very small study, presenting data from just 16 women.

* Note that this study was published as an abstract and presented at a conference. These data and conclusions should be considered to be preliminary until published in a peer-reviewed journal.

SAN DIEGO -- Drinking too many diet soft drinks may result in a negative calcium balance, a marker of low bone mineral density, researchers here reported.

Mean calcium excretion over three hours after drinking diet cola was 6.85 mg higher than after drinking water (P=0.004), Noelle Larson, MD, of the Walter Reed Army Medical Center, in Washington, and colleagues reported at the annual meeting of the Endocrine Society.

In addition, mean phosphorous excretion was 41 mg higher in the cola group (P=0.003).

Larson said that she became interested in the topic after seeing responses on informal surveys of young women medical students some of whom reported drinking 20 to 24 Diet Cokes a week.

"Just observationally as a young woman with friends, I see that people who drink diet sodas tend to drink them in a different pattern compared to people who drink regular sodas, as a chronic, repeated thing," often in an effort to lose weight or prevent weight gain, Larson said.

She noted that several cross-sectional studies show cola beverages are associated with increased fracture risk and decreased bone mineral density.

In an earlier investigation, the researchers had looked at the hormonal effects of diet cola ingestion on parathyroid hormone, calcium, phosphorus, insulin, alkaline phosphotase, and ghrelin.

The researchers thought that because of the phosphorus load, PTH would surge, but they found exactly the opposite, "which was that it comes down and sort of comes back to baseline; alkaline phosphatase increases also," Larson said. "We thought, 'Well, that suggests there's some turnover of bone going on, and maybe there's some calcium being mobilized and it's going out in the urine and that might partially account for the fracture risk and decreased bone density that's being described."

With results from that earlier study as the impetus, Larson and colleagues undertook the current study, for which they recruited 20 healthy women, ages 18 to 40.

Exclusion criteria were fracture within the prior six months, known bone disease or vitamin D deficiency, steroid or diuretic use, breast-feeding, and vitamin D supplementation above the current U.S. recommended daily allowance.

The participants were randomized to drink 24 ounces of either water or diet cola on two study days. Urine was collected for three hours after ingestion of the designated beverage and assayed for calcium, phosphorous, and creatinine using standard assays.

Data were analyzed on 16 participants; four were excluded because of lab error or failure to comply with the study protocol, the researchers said.

In addition to the higher calcium and phosphorus excretion, the investigators also found that normalized calcium and phosphorous excretion per gram of creatinine showed a trend in the same direction as total calcium and phosphorous per three hours. That figure did not achieve statistical significance, however.

Although the study was small, "it does look like there was a statistically significant rise in urine calcium," said Larson. "The important part about that is that Diet Coke has no calcium content."

Compared with milk, which also causes a rise in urine calcium but is replacing calcium at the same time, diet colas "would [create] an overall negative body calcium balance and that could partially explain why they appear to be bad for bones," she said.

Although the study is too small to draw any firm conclusions, "certainly my personal practice among adolescent girls who tend to be concerned about their weight -- and who drink diet beverages while they are in that critical period of bone formation -- is to just try and counsel them to set habits of drinking calcium-containing beverages and maintaining adequate vitamin D," said Larson.

Elizabeth Barrett-Connor, MD, of the University of California San Diego, called the study "fabulous."

Barrett-Connor, who was not involved in the study, said that although it was a small and short-term trial, "it fits with all my preconceived ideas" about the nutritional problems with diet soda. "This [is new] but it just makes sense."

The study was funded by the Walter Reed Department of Clinical Investigations.

* Note that this study was published as an abstract and presented at a conference. These data and conclusions should be considered to be preliminary until published in a peer-reviewed journal.

SAN DIEGO -- Males presenting with high athletic stress or weight loss, coupled with low testosterone, may signal the rise of a new disorder -- functional hypogonadotrophic hypogonadism.

Just as women whose bodies are under stress from excessive exercise, weight loss, or psychological stress can experience hypothalamic amenorrhea, a seven-patient series suggests that a similar phenomenon may exist among men undergoing similar kinds of stress, Andrew Dwyer, MD, of Massachusetts General Hospital, said during a poster session here at the annual meeting of the Endocrine Society.

"We saw some male patients who all have a similar type of presentation in terms of one or more of this [stress] triad, and presented with low testosterone," Dwyer explained. The patients all had normal puberty and a normal testicular size, but all presented with "vague, non-specific symptoms" of low testosterone, including absent morning erections, low energy level, fatigue, decreased athletic performance, and decreased libido, he said.

"Interestingly, two of these patients had female family members with amenorrhea, which made us think maybe there's a connection," Dwyer continued.

To further study this phenomenon, the patients were recalled to the hospital, where they underwent detailed genotyping and phenotyping, including measurements of reproductive and metabolic hormones, an overnight frequent sampling study of leutinizing hormone, and DEXA scan for body composition. The investigators also recruited 35 age-matched healthy adults as controls.

The seven patients had a lower average weight compared to controls (64.1 kg versus 79.9 kg, P<0.01). They also had a lower body mass index (20.7 versus 24.9, P<0.01) and a lower percentage of body fat (9.8% versus 17.6%, P<0.01).

The patients also had lower pulse frequency, lower mean LH amplitude, and lower serum FSH, but none of those numbers approached statistical significance, according to the investigators.

Despite their low testosterone levels, six of the seven patients had LH pulse patterns, frequency, and amplitude that were no different from controls, Dwyer said. However, the seventh patient had four hours of no pulses, then a burst of three pulses, then no pulses for the remaining four hours, a pattern that normally occurs when boys first enter puberty.

"It's as if this patient is recapitulating an early- to mid-pubertal LH pulse secretion pattern," he said. "He's 17, he went through normal puberty, he's done and he's virilized, but with the stress of exercise and the weight loss, perhaps the stress tipped him back into the nocturnal pulse pattern."

Dwyer noted that after the patients had been tested, one of them sustained a heel injury and had to stop training for a while. "He gained six pounds, and we measured his testosterone level, and serially, it stayed normal," he noted. "So with just enough removal of stress...he was able to swing back into normal testosterone production."

Another patient who decreased his training upon the researchers' recommendation was also retested and his testosterone level was up into the low end of the normal range, said Dwyer.

The researchers are calling the possible new disorder functional hypogonadotrophic hypogonadism. "In Boston, there are lots of marathon runners and collegiate rowers who exercise a lot and don't exhibit these symptoms," he said. "So what is it about these seven men that make them different from vast majority of superexerciser lean guys?"

The investigators hypothesize that these men may harbor mutations in genes that are involved in GnRH androgyny or reproductive access such that with the right stressor, that can tip them into hypogonadism, but if you remove the stressor they tip back," said Dwyer.

Rick Dorin, MD, chief of endocrinology at the University of New Mexico, in Albuquerque, said the study was very interesting.

"I see a painfully large amount of hypogonadism in in my clinical practice at the Veterans Affairs Hospital," said Dorin, who was not involved in the study. "We see a lot of hypogonadism due to other factors, but not in such young men. This is raising the possibility that the [hypothalamic amenorrhea] in women athletes -- that a comparable thing goes on in young men. They've got provocative findings in a small number of patients."

The study was funded by the National Institutes of Health. Dwyer and Dorin each reported having no conflicts of interest.

Health Starts with LifestyleConcerned with the rising cost of health Care?

Every wonder why medication has more side effects then benefits?

Have you every been curious about alternative methods for health?

If you answered yes to these questions, you may find what I'm about to tell you interesting. For the last two years I have researched alternative methods for improving health with the absence of costly medications and doctors visits. I have personally found the results of my research to be highly beneficial.

SOTA Instruments, has perfected a little known technology. Originally discovered by researches at the Albert Einstein College of Medicine, and then rediscovered by Dr. Robert Beck. This technology has profound and patented health benefits, through the utilization of electronic micro currents. Legally issued U.S. patents hold facts which may shock you.

Below is an informative video of Dr. Bob Beck. His information WILL shock you!

Health Starts with LifestyleConcerned with the rising cost of health Care?

Every wonder why medication has more side effects then benefits?

Have you every been curious about alternative methods for health?

If you answered yes to these questions, you may find what I'm about to tell you interesting. For the last two years I have researched alternative methods for improving health with the absence of costly medications and doctors visits. I have personally found the results of my research to be highly beneficial.

SOTA Instruments, has perfected a little known technology. Originally discovered by researches at the Albert Einstein College of Medicine, and then rediscovered by Dr. Robert Beck. This technology has profound and patented health benefits, through the utilization of electronic micro currents. Legally issued U.S. patents hold facts which may shock you.

Below is an informative video of Dr. Bob Beck. His information WILL shock you!

Preclinical Animal Data Demonstrates Promising Stem Cell Therapeutic Strategy for HIV/AIDS - Extends ZFP Therapeutic ApplicationRICHMOND, Calif., July 6 /PRNewswire-FirstCall/ -- Sangamo BioSciences, Inc. (Nasdaq: SGMO) announced the publication of data demonstrating the preclinical efficacy of a human stem cell therapy for human immunodeficiency virus (HIV) based on its proprietary zinc finger DNA-binding protein nuclease (ZFN) technology. The ZFN approach enables the permanent disruption of the CCR5 gene, which encodes an important receptor for HIV infection, in all the cell types comprising the immune system that develop from hematopoietic stem cells (HSCs), and is the basis for a promising therapeutic strategy for the treatment of HIV/AIDS. Sangamo has two ongoing Phase 1 clinical trials to evaluate the safety and clinical efficacy of this approach in CD4+ T-cells.

The work, which was carried out in the laboratory of Paula Cannon, Ph.D., Associate Professor of Molecular Microbiology & Immunology at the Keck School of Medicine of the University of Southern California (USC), in collaboration with Sangamo scientists, was published on July 2, 2010, as an Advance Online Publication (http://www.nature.com/nbt/journal/vaop/ncurrent/full/nbt.1663.html) in Nature Biotechnology.

"These are very exciting data that provide proof of concept for a new approach to HIV treatment," said John Zaia, M.D., the Aaron D. and Edith Miller Chair in Gene Therapy and Chair of Virology, City of Hope. "The recent example of the 'Berlin Patient' who appears to have been cured of both his HIV and leukemia by receiving a bone marrow transplant (BMT) of stem cells from a donor that had a naturally occurring CCR5 mutation that makes them resistant to HIV infection, provided the model for this approach. However, the paucity of human donors with this natural CCR5 mutation and the risks of allogeneic BMT mean that we need a more practical solution to make this a therapeutic option. Modification of HSCs using ZFNs to recreate the CCR5 mutation is a potential solution."

Dr. Zaia is the leader of the recent $14.5 million Disease Team Research Award granted by the California Institute for Regenerative Medicine (CIRM) to a multidisciplinary team of investigators which includes City of Hope, Dr. Cannon and her colleagues at USC, and Sangamo scientists. The award funds the preclinical development of a ZFN CCR5-targeted approach which aims to complete an Investigational New Drug (IND) application to the U.S. Food and Drug Administration (FDA) for clinical testing of this ZFN method.

Sangamo's ZFNs are designed to permanently modify the DNA sequence encoding CCR5, a co-receptor that enables HIV to enter and infect cells of the immune system. Individuals carrying a naturally occurring mutation of their CCR5 gene, a variant known as CCR5-delta32, have been shown to be resistant to HIV infection. Building on this observation, a study published in the New England Journal of Medicine in 2009 reported a potential "cure" when an AIDS patient with leukemia received a bone marrow transplant from a "matched" donor with this delta-32 CCR5 mutation. This approach transferred the HSCs residing in the bone marrow from the delta-32 donor, and provided a self-renewable and potentially lifelong source of HIV-resistant immune cells. After transplantation, the patient was able to discontinue all anti-HIV drug treatments, CD4 counts increased, and viral load dropped to an undetectable level, demonstrating effective transplantation of protection from HIV infection. The data reported in the Nature Biotechnology publication replicate these findings for a ZFN-based treatment in a preclinical model.

"The data described in this paper are an important demonstration of the potential therapeutic possibilities of ZFN modification of human stem cells," commented Philip Gregory, D. Phil., Sangamo's vice president of research and chief scientific officer. "We have demonstrated efficient and specific modification of human hematopoietic stem cells, rendering them resistant to infection with HIV-1 while retaining their 'stemness' and ability to differentiate. These data pave the way for the use of this technology in other diseases for which HSC modification may be therapeutically useful."

Data Reported in the Nature Biotechnology Paper

The reported results demonstrate that a one-time exposure to CCR5-specific ZFNs resulted in the generation of an HIV-resistant population of human HSCs by the permanent genetic modification of the CCR5 gene. These ZFN-modified stem cells engrafted in NSG (NOD/SCID/IL2rγnull) mice, which lack a normal immune system and are able to tolerate engraftment of human cells and tissues. After 8-12 weeks the engrafted ZFN-modified human cells could be identified as different immune cell types in the peripheral blood, and various tissues of the mouse suggesting that they were functionally normal. Furthermore, the ZFN-modified HSCs produced progeny that could be harvested from one mouse and engrafted into a second animal, demonstrating that the modified HSCs retain their 'stemness' and ability to differentiate. In addition, the animals did not experience any obvious toxicity or ill-health. In HIV challenge experiments, researchers found that the ZFN-modified cells had a selective advantage over unmodified HSCs and not only survived infection but expanded and appeared to traffic normally to various tissues in the mouse. Moreover, the presence of ZFN-modified cells controlled HIV replication in the animals. These data suggest that human HSCs can be modified with ZFNs, expand and differentiate and have a selective advantage in the presence of HIV allowing them to evade infection and destruction leaving them able fight opportunistic infections and HIV itself.

About HIV/AIDS and CCR5

Human Immunodeficiency Virus (HIV) infection kills or impairs cells of the immune system, progressively destroying the body's ability to fight infections and certain cancers resulting in AIDS (Acquired Immune Deficiency Syndrome). Individuals diagnosed with AIDS are susceptible to life-threatening diseases called opportunistic infections, which are caused by microbes that usually do not cause illness in healthy people. According to UNAIDS/WHO, over 2.7 million people were newly infected with HIV in 2007. An estimated 2.0 million people died of AIDS in the same year. There are now over 33 million people living with HIV and AIDS worldwide. The CDC estimates that, in the United States alone, there were 1.2 million people living with HIV/AIDS, approximately 54,000 new infections and 23,000 deaths in 2007.

CCR5 is the chemokine receptor that HIV uses as a co-receptor to gain entry into immune cells. CCR5 is perhaps the most important of the known co-receptors for HIV, since the most commonly transmitted strains of HIV are strains that bind to CCR5 -- so-called "R5" strains. A small fraction of the population carries a mutation in their CCR5 gene, called the delta32 mutation. This mutated version of the gene results in a truncated CCR5 protein which cannot be used by HIV as a co-receptor. Individuals that have mutant delta 32 versions of both of their CCR5 genes are resistant to infection by R5 HIV strains.

About Sangamo

Sangamo BioSciences, Inc. is focused on the research and development of novel DNA-binding proteins for therapeutic gene regulation and modification. The most advanced ZFP Therapeutic™ development program is currently in a Phase 2b clinical trial for evaluation of safety and clinical effect in patients with diabetic neuropathy and a Phase 2 trial in ALS. Sangamo also has two Phase 1 clinical trials to evaluate safety and clinical effect of a treatment for HIV/AIDS and another Phase 1 trial to evaluate safety and clinical effect of a treatment for recurrent glioblastoma multiforme. Other therapeutic development programs are focused on neuropathic pain, nerve regeneration, Parkinson's disease and monogenic diseases. Sangamo's core competencies enable the engineering of a class of DNA-binding proteins known as zinc finger DNA-binding proteins (ZFPs). By engineering ZFPs that recognize a specific DNA sequence Sangamo has created ZFP transcription factors (ZFP TF) that can control gene expression and, consequently, cell function. Sangamo is also developing sequence-specific ZFP Nucleases (ZFN) for gene modification. Sangamo has established strategic partnerships with companies in non-therapeutic applications of its technology including Dow AgroSciences and Sigma-Aldrich Corporation. For more information about Sangamo, visit the company's website at http://www.sangamo.com/.

This press release may contain forward-looking statements based on Sangamo's current expectations. These forward-looking statements include, without limitation, references to the research and development of novel ZFP TFs and ZFNs as ZFP Therapeutics, applications of Sangamo's ZFP technology platform, the therapeutic potential of ZFNs for the treatment of HIV/AIDS, strategic partnerships with collaborators and clinical trials of ZFP Therapeutics. Actual results may differ materially from these forward-looking statements due to a number of factors, including technological challenges, uncertainties relating to the initiation and completion of stages of ZFP Therapeutic clinical trials, Sangamo's ability to develop commercially viable products and technological developments by our competitors. See the company's SEC filings, and in particular, the risk factors described in the Sangamo's Annual Report on Form 10-K and its most recent Quarterly Report on Form 10-Q. Sangamo BioSciences, Inc. assumes no obligation to update the forward-looking information contained in this press release.

Scientists have discovered two potent human antibodies that can stop more than 90 percent of known global HIV strains from infecting human cells in the laboratory, and have demonstrated how one of these disease-fighting proteins accomplishes this feat. According to the scientists, these antibodies could be used to design improved HIV vaccines, or could be further developed to prevent or treat HIV infection. Moreover, the method used to find these antibodies could be applied to isolate therapeutic antibodies for other infectious diseases as well.

“The discovery of these exceptionally broadly neutralizing antibodies to HIV and the structural analysis that explains how they work are exciting advances that will accelerate our efforts to find a preventive HIV vaccine for global use,” says Anthony S. Fauci, M.D., director of the National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health. “In addition, the technique the teams used to find the new antibodies represents a novel strategy that could be applied to vaccine design for many other infectious diseases.”

Led by a team from the NIAID Vaccine Research Center (VRC), the scientists found two naturally occurring, powerful antibodies called VRC01 and VRC02 in an HIV-infected individual's blood. They found the antibodies using a novel molecular device they developed that homes in on the specific cells that make antibodies against HIV. The device is an HIV protein that the scientists modified so it would react only with antibodies specific to the site where the virus binds to cells it infects.

The scientists found that VRC01 and VRC02 neutralize more HIV strains with greater overall strength than previously known antibodies to the virus.

The researchers also determined the atomic-level structure of VRC01 when it is attaching to HIV. This has enabled the team to define how the antibody works and to precisely locate where it attaches to the virus. With this knowledge, they have begun to design components of a candidate vaccine that could teach the human immune system to make antibodies similar to VRC01 that might prevent infection by the vast majority of HIV strains worldwide.

NIAID scientists Peter D. Kwong, Ph.D., John R. Mascola, M.D., and Gary J. Nabel, M.D., Ph.D., led the two research teams. A pair of articles about these findings appears today in the online edition of Science.

"We have used our knowledge of the structure of a virus — in this case, the outer surface of HIV — to refine molecular tools that pinpoint the vulnerable spot on the virus and guide us to antibodies that attach to this spot, blocking the virus from infecting cells," explains Dr. Nabel, the VRC director.

Finding individual antibodies that can neutralize HIV strains anywhere in the world has been difficult because the virus continuously changes its surface proteins to evade recognition by the immune system. As a consequence of these changes, an enormous number of HIV variants exist worldwide. Even so, scientists have identified a few areas on HIV's surface that remain nearly constant across all variants. One such area, located on the surface spikes used by HIV to attach to immune system cells and infect them, is called the CD4 binding site. VRC01 and VRC02 block HIV infection by attaching to the CD4 binding site, preventing the virus from latching onto immune cells.

"The antibodies attach to a virtually unchanging part of the virus, and this explains why they can neutralize such an extraordinary range of HIV strains," says Dr. Mascola, the deputy director of the VRC.

With these antibodies in hand, a team led by Dr. Kwong, chief of the structural biology section at the VRC, determined the atomic-level molecular structure of VRC01 when attached to the CD4 binding site. They then examined this structure in light of natural antibody development to ascertain the steps that would be needed to elicit a VRC01-like antibody through vaccination.

Antibody development begins with the mixing of genes into new combinations within the immune cells that make antibodies. Examination of the structure of VRC01 attached to HIV suggested that, from a genetic standpoint, the immune system likely could produce VRC01 precursors readily. The researchers also confirmed that VRC01 does not bind to human cells — a characteristic that might otherwise lead to its elimination during immune development, a natural mechanism the body employs to prevent autoimmune disease.

Image of the Atomic structure of the antibody VRC01 binding to HIV.

Atomic structure of the antibody VRC01 (blue and green) binding to HIV (grey and red). The precise site of VRC01-HIV binding (red) is a subset of the area of viral attachment to the primary immune cells HIV infects. Credit: NIAID VRC

In the final stage of antibody development, antibody-producing B cells recognize specific parts of a pathogen and then mutate, or mature, so the antibody can bind to the pathogen more firmly. VRC01 precursors do not bind tightly to HIV, but rather mature extensively into more powerfully neutralizing forms. This extensive antibody maturation presents a challenge for vaccine design. In their paper, Dr. Kwong and colleagues explore how this challenge might be addressed by designing vaccine components that could guide the immune system through this stepwise maturation process and facilitate the generation of a VRC01-like antibody from its precursors. The scientists currently are performing research to identify these components.

"The discoveries we have made may overcome the limitations that have long stymied antibody-based HIV vaccine design," says Dr. Kwong.

The two research teams included NIAID scientists from the VRC, the Laboratory of Immunoregulation, and the Division of Clinical Research, all in Bethesda, Md.; as well as researchers from Beth Israel Deaconess Medical Center in Boston; Columbia University in New York; Harvard Medical School and Harvard School of Public Health in Boston; The Rockefeller University in New York City; and University of Washington in Seattle.

NIAID conducts and supports research — at NIH, throughout the United States, and worldwide — to study the causes of infectious and immune-mediated diseases, and to develop better means of preventing, diagnosing and treating these illnesses. News releases, fact sheets and other NIAID-related materials are available on the NIAID Web site at http://www.niaid.nih.gov.

The National Institutes of Health (NIH) — The Nation's Medical Research Agency — includes 27 Institutes and Centers and is a component of the U.S. Department of Health and Human Services. It is the primary federal agency for conducting and supporting basic, clinical and translational medical research, and it investigates the causes, treatments, and cures for both common and rare diseases. For more information about NIH and its programs, visit www.nih.gov.

Now the next step for them is to try the research on humans; they hope that it'll work for us. If it works, patients don't need to take anti-retroviral medicines anymore all they have to do is undergo stem cell therapy to create HIV resistant immune cells that can kill the HIV viruses in their body."In the presence of CCR5-tropic HIV-1, CCR5-/- progeny rapidly replaced cells depleted by the virus, leading to a polyclonal population that ultimately preserved human immune cells in multiple tissues. Our findings indicate that the modification of only a minority of human CD34+ HSPCs may provide the same strong anti-viral benefit as was conferred by a complete CCR5Δ32 stem cell transplantation in a patient9. And they further suggest that a partially modified autologous transplant, administered under only mildly ablative transplantation regimens may also be effective, opening up the treatment to many more HIV-infected individuals. Finally, the identification of conditions that allow the efficient use of ZFNs in human CD34+ HSPCs suggests the use of this technology in other diseases for which HSPC modification may be curative."

"mice transplanted with ZFN-modified HSPCs underwent rapid selection for CCR5-/- cells, had significantly lower HIV-1 levels and preserved human cells throughout their tissues. The demonstration that a minority of CCR5-/- HSPCs can populate an infected animal with HIV-1-resistant, CCR5-/- progeny supports the use of ZFN-modified autologous hematopoietic stem cells as a clinical approach to treating HIV-1."

"An alternative approach to controlling HIV-1 replication is engineering of the body's immune cells to be resistant to infection44. In this regard, the CCR5 co-receptor is an attractive target because of the HIV-resistant phenotype of homozygous CCR5Δ32 individuals3. In the present study, we identified conditions that allow efficient disruption of CCR5 in human CD34+ HSPCs and demonstrated that such modified cells generate CCR5-/-, HIV-resistant progeny in a mouse model of human hematopoiesis and HIV-1 infection, leading to control of HIV-1 replication.."

HIV-resistant cells work in mice. Can they help humans?California scientists, boosted by stem cell research funding enabled by Proposition 71, are aiming for clinical trials involving gene therapy through bone marrow transplants.

Paula Cannon, a biology professor at USC's Keck School of Medicine, inspects a mouse that will be infected with HIV. (Allen J. Schaben, Los Angeles Times / July 16, 2010)

The stink came from scores of little white mice scurrying about in cages. Some of the cages were marked with red biohazard signs, indicating mice that had been injected with HIV.

Yet, in some of the animals — ones with a small genetic change — the virus never took hold.

Like mouse, like man? Maybe so.

In early 2007, a patient in Berlin needed a bone marrow transplant to treat his leukemia. He was also HIV positive, and his doctor had an idea: Why not use the marrow from one of the rare individuals who are naturally resistant to HIV and try to eradicate both diseases at once?

It worked. Sixty-one days after the patient's transplant, his virus levels were undetectable, and they've stayed that way.

Since news of the man's cure broke, HIV patients have been telephoning doctors to ask for bone marrow transplants. But it's not that simple. The treatment is too risky and impractical for widespread use.

"A bone marrow transplant — it's a horrible process you would not wish on your worst enemy unless they needed one to save their life," said Cannon, a biology professor at USC's Keck School of Medicine. There are grueling treatments to prepare a patient for transplant; the danger of rejecting the marrow; and the risk of graft-versus-host disease, wherein the marrow attacks the patient.

And that's assuming the patient can find a matching donor — a difficult proposition in itself — with the right HIV-resistant genetic constitution, which is present in only about 1% of the Caucasian population.

But there could be another way.

Instead of sifting through the sands for a rare donor and then subjecting a patient to the dangers of a bone marrow transplant, Cannon and her colleague Philip Gregory, chief scientific officer at the Richmond, Calif.-based biotech company Sangamo BioSciences, began to think: They could use gene therapy instead, to tweak a patient's own cells to resistance — and recovery.

The mouse "cure," they say, suggests they're on the right track.

Now, with $14.5 million from the California Institute for Regenerative Medicine, the San Francisco-based stem cell research-funding center created by 2004's Proposition 71, Cannon, Gregory and researchers at the City of Hope cancer center in Duarte are working toward bringing the technique to clinical trials within four years.

Cannon and other HIV researchers insist that, despite cancers and deaths associated with past gene therapy trials, it's the right way to target the disease. They cite recent successes, including treatments that cured children with the "bubble boy" syndrome and helped blind children regain their vision.

"I don't think anyone would want to do gene therapy if there were an alternative," said Caltech biologist David Baltimore, one of the many L.A.-based researchers pursuing gene therapy strategies to prevent or cure HIV. "I think it's absolutely necessary. Nothing else will work."

Since AIDS emerged in the early 1980s, development of anti-HIV medications has turned the disease from a virtual death sentence into a chronic, manageable condition.

But the clamor for a cure hasn't quieted.

Vaccine trials have failed; drug-resistant strains are on the rise; and the meds, which can have uncomfortable side effects such as fatigue, nausea and redistribution of body fat that creates a so-called buffalo hump, cost about $20,000 a year.

A bone marrow transplant is about five times as expensive, but it would have to be done only once.

The question was, could researchers create bone marrow stem cells that — just like the marrow the Berlin patient received — lack the crucial gene, CCR5, that normally lets HIV into the key immune cells it destroys?

In 2006, Gregory asked Cannon if she was interested in testing whether a tool his company developed, called a zinc finger nuclease, could do the trick.

Zinc finger nucleases are genetic scissors, cutting DNA at a specific site — say, in the middle of the CCR5 gene. When the cell glues the gene back together, it usually makes a mistake, resulting in a gene that no longer works.

The team spent about a year optimizing the procedure for treating delicate stem cells with the CCR5 snippers.

They tested the method using so-called humanized mice — ones engineered to have a human immune system — because HIV doesn't infect normal mice. When stem cells were treated with the molecular scissors before being injected into mice, the resulting immune system lacked CCR5, exactly as the scientists had hoped.

These mice acted just like the Berlin patient — they fought off the virus.

Ready to make the leap from mouse to man, Gregory found a third leg for the team: researchers at City of Hope, who had extensive bone marrow transplant expertise.

"They brought Paula's data to us and we said, 'Wow, this looks fantastic,'" said Dr. John Zaia, City of Hope's deputy director for clinical research.

Researchers there are now working toward clinical trials, optimizing every element of the treatment for safety, effectiveness and reproducibility.

On a wiltingly hot afternoon in July, lab manager Lucy Brown maneuvered a computer mouse across three screens speckled with red, yellow and green dots.

The computer was hooked to a flow cytometer — a collection of black boxes, green wires and silver knobs that can detect subtle differences between cells and separate them at a rate of 50,000 per second. This is how the scientists will separate stem cells from patients' blood once trials are underway, to be sure that the genetic fix in the CCR5 gene was made, and kept.

Upstairs, machines with mazes of sterile tubes and pumps stood ready to prepare cells for CCR5-snipping. Here, the scientists will purify the bone marrow stem cells, increasing their numbers first to 5% of total cells, up from a measly 0.1% in the starting mixture, and then to 99%. At this point they can begin testing methods to clip the cells' DNA.

When all is perfected, the scientists will have a precise recipe for producing batches of engineered stem cells, including exactly how long the cells should be treated, how much of each chemical needs to be added, how pure the cells need to be, and thousands of other details.

"We are literally writing the book on how you do this," said David DiGiusto, director of City of Hope's bone marrow stem cell therapy research.

To receive FDA approval for clinical trials — a goal they hope to achieve in three to four years — the researchers must prove that they can safely and reliably prepare the cells. Once they get the green light, the first cases will probably be people like the Berlin patient who need bone marrow transplants to treat AIDS-related lymphoma.

They'll modify the patients' cells in the stringently sterile manufacturing lab that DiGiusto designed with details such as cove molding and seamless floors so there are no corners or cracks to collect dust. Anyone who enters must wear a full bunny suit, much like the one Cannon wears in her mouse room, to keep from contaminating the delicate cells.

Some have advertised the effort as a quest for the elusive "C" word, but Cannon doesn't quite see it that way.

"People say we're trying to cure HIV," she said. "I think of it more as, we're just trying to make the body live quite happily and healthily with a small amount of virus."

"I don't think anyone would want to do gene therapy if there were an alternative," said Caltech biologist David Baltimore, one of the many L.A.-based researchers pursuing gene therapy strategies to prevent or cure HIV. "I think it's absolutely necessary. Nothing else will work."

Researchers at Loyola University Health Systems announced an important breakthrough Aug. 20, one that could lead to a treatment that destroys the HIV virus.

Senior researcher Edward Campbell (photo below) and his colleagues reported that they’ve identified key components of a protein called TRIM5a that kills the virus in rhesus monkeys. The finding could lead to TRIM5a-based treatments for people that would "knock out" the HIV virus.

The Loyola team started studying TRIM5a after other researchers reported in 2004 that it protects rhesus monkeys from HIV by first latching onto the virus; other TRIM5a proteins then "gang up" on the virus and destroy it, the researchers found.

Humans also carry the protein, but while it protects people from some viruses it does not protect them from HIV. So the Loyola team looked for components of TRIM5a that kill HIV in rhesus monkeys. By doing that, they hope to turn TRIM5a into an effective weapon against HIV in humans.

"Scientists have been trying to develop antiviral therapies for only about 75 years," Campbell said. "Evolution has been playing this game for millions of years, and it has identified a point of intervention that we still know very little about."

TRIM5a consists of almost 500 amino acid sub-units. The Loyola team has identified six that play a critical role in the protein’s ability to fight viral infection. When those six amino acids were altered in human cultures, the protein lost its ability to block HIV infection, the researchers said.

The research may also lead to the development of drugs that mimic TRIM5a’s actions against HIV. The team uses Loyola’s wide-field "deconvolution" microscope to study TRIM5a.

"The motto of our lab is one of Yogi Berra’s sayings - ’You can see a lot just by looking,’" Campbell said.

The team’s findings are featured in a cover story in the Sept. 15 issue of the journal Virology, now available online. In addition to Campbell, assistant professor in the Department of Microbiology and Immunology at Loyola University Chicago Stritch School of Medicine, the co-authors of the study are Jaya Sastri, a Stritch graduate student; Christopher O’Connor, a former post-doctorate researcher at Stritch; Cindy Danielson and Michael McRaven, of Northwestern University Feinberg School of Medicine; and Patricio Perez and Felipe Diaz-Griffero, of Albert Einstein College of Medicine.

* Explain to interested patients that a series of abstracts suggests that marathon-type running does not appear to induce long-term changes in the heart, even in older people.

* Note that one researcher suggests that novice older runners should still undergo basic heart tests before training for marathons or ultramarathons.

* Note that these studies were published as abstracts and/or presented at a conference. These data and conclusions should be considered to be preliminary until published in a peer-reviewed journal.

STOCKHOLM -- Men and women who participate in endurance competitive marathon events appear to develop some transient heart changes, but overall these activities do not seem to have long-term harm for the vast majority of individuals.

In a series of reports presented at the European Society of Cardiology meeting, doctors found:

* Older runners -- those over age 50 -- developed some cardiac changes following running in Berlin marathons, but the changes in diastolic and right heart function did not exceed normal ranges.

* There are ethnic and sexual differences in changes in heart muscle that should be recognized before athletes with enlarged hearts are disqualified from competition.

* No significant cardiac changes occurred among participants who were engaged in a week-long overland and water endurance exercise program.

* On the other hand, ultra-endurance running -- races of 50 to 100 miles -- resulted in elevation of troponin-1 which could be related to heart muscle damage; many of these runners also developed electrocardiographic changes.

On the whole, "It is wonderful to see that older adults can participate in these endurance events without experiencing long-term heart damage," said Ileana L. Pina, MD, professor of medicine, epidemiology, and biostatistics at Case Western Reserve University, in Cleveland.

"Even if these individuals exhibit higher-than-normal biomarkers, these rapidly return to normal," Pina, a spokesperson for the American Heart Association, told MedPage Today in Stockholm.

In one study, researchers led by Fabian Knebel, MD, a cardiologist at the Medical Clinic for Cardiology, Angiology, Pneumology at the University Medicine Berlin, studied cardiac parameters for 167 runners -- all 50 years or older -- who participated in the 2006 and 2007 Berlin Marathon races.

They were examined 10 days before the race, immediately after running 26 miles, 385 yards in the marathon, and two weeks after the race.

Knebel said that immediately after the race, the heart rate in these runner was elevated from baseline -- 88 beats per minute at the end of the marathon compared with 62 beats per minute before the race (P<0.001), and 57.95 of the runners showed increases in troponin values.

However, these changes proved transient, he said. "Two weeks after a marathon, the key parameters were all back to normal levels," Knebel said. "The concerns people have about marathon running causing sustained damage to the heart appear to be unfounded."

In another study, researchers determined that Caucasian and black women athletes appear to have differences in anatomy which manifests as a larger heart among the black women.

Sanjay Sharma, MD, professor of cardiology at Kings College, in London, established that about 3% of black women athletes have a left ventricular wall thickness greater than 11 mm -- a level not exceeded by elite white women athletes. In black elite women athletes, the left ventricular wall thickness can reach 13 mm -- a level that would be considered abnormal for white women athletes.

He suggested that if sports policymakers used data derived solely from white athletes regarding what constitutes healthy heart function, "it could unfairly discriminate against black athletes by leading to unnecessary investigation or even disqualification."

In commenting on Sharma's study, Pina noted, "We have known there are differences between the sexes in cardiology, but it also appears there are genetic differences that involve ethnicity as well."

A third study that examined subjects who completed a five to seven day prolonged exercise event -- an 800 kilometer cross-country running and boating test -- found that the athletes who participated experienced no health-related abnormalities.

Researcher C. Mikael Mattsson, PhD, at the Karolinska Institute, Stockholm, suggested that the lack of cardiac fatigue seen among the 15 athletes studied might have been due to the low intensity exercise of the cross-country trek. "This might point towards exercise intensity, not duration, as the primary source for cardiac fatigue," he said.

That might have been borne out among participants who completed either a 50-mile or a 100-mile road race, said John Somauroo, MD, professor of medicine at Countess of Chester Hospital, England.

Of the 25 runners who completed the arduous race -- beset by thunderstorms and driving rain -- troponin levels were elevated in 21 of the men, and many developed bizarre electrocardiogram changes, Sumauroo and colleagues found.

"This study suggests that running continuously over 50 to 100 miles may not be good for the heart," Somauroo said.

Aside from the long-distance studies, the lack of long-term damage to these athletes' hearts may point to the human genetic makeup. "In evolution we were selected out as hunter-gatherers where we would run and hunt miles from home to find enough food for the day," Stephen Gielen, MD, associate professor of medicine at the University of Leipzig, Germany, and a spokesman for the European Society of Cardiology, told MedPage Today.

"It is astonishing the enormous exercise that the human heart can endure," he said. He noted that the long-distance running may be typical of the human condition that was developed before mankind became civilized.

He did note that the long-distance running events that Somauroo reported may represent the "edge of harmful" events.

Gielen said that individuals who decide to take up endurance running and are over age 40 should undergo cardiovascular screening before hitting the road.

from jules: As I have reported previously we know muscle wasting occurred for many HIV-infected in the earlier years back in the 80s and 90s, and bone loss and fat loss (lipoatrophy) is a concern for many with HIV. Bone and muscle loss are related, so it appears that individuals HIV-infected years ago that have suffered muscle, bone & fat loss may be facing aging problems. I think that mitochondrial loss which many with HIV have suffered may contribute to muscle loss, as well diabetes & insulin resistance may as well. The MACS has reported HIV+ experience frailty 10 years earlier than HIV-negatives. This is why the NIH should declare aging/HIV an emergency, its not enough just to conduct research, we need to address these concerns in an expedited manner. Exercise is considered the way to prevent muscle loss.

“In the future, sarcopenia will be known as much as osteoporosis is now,” said Dr. Bruno Vellas, president of the International Association of Gerontology and Geriatrics......Experts say the best approach to restoring or maintaining muscle mass and strength is exercise, particularly resistance training.......Maintaining the muscle is possible,” said Dr. Luigi Ferrucci (will be speaking at Aging/HIV Workshop in October) of the National Institute on Aging, who directs the study, called the Baltimore Longitudinal Study of Aging. “We just don?t know the right formula yet......The National Institute on Aging is now sponsoring a controlled trial to test whether exercise can prevent disability in largely sedentary people, age 70 to 89. There is also some early evidence that nutrition, like vitamin D or high levels of protein, might help. “At this point, what we can say is that older people are at risk for eating too little protein for adequate muscle preservation,” said Dr. Elena Volpi of the University of Texas Medical Branch in Galveston......Pharmaceutical companies are paying more attention to muscles, a part of the body they once largely ignored. A year ago, for instance, GlaxoSmithKline hired William Evans, a leading academic expert on sarcopenia, to run a new muscle research unit......But some studies have shown that strength, such as gripping force, or muscle function, as measured, say, by walking speed, (frailty) are more important than mass in predicting problems seniors might have......Efforts to develop muscle drugs are still in early stages, and there have been setbacks.

"Aging, even in healthy elderly people, is accompanied by a reduction in muscle mass and muscle strength....Vitamin D deficiency is associated with muscle weakness....Because 1,25(OH)D3 exerts its influence on distant target tissue, mediated by a vitamin D receptor (VDR), it is considered to be a hormone rather than a vitamin....Supplementation in this population improved muscle strength, walking distance, functional ability (57-59), and body sway (70). These findings and the observed improvements in bone density after vitamin D supplementation (67, 72) provide an explanation for the association between vitamin D supplementation and fewer falls and nonvertebral fractures in elderly people (69, 71)."

GSK signs Muscle Drug Deal, $500 million cancer pact with AmplimmuneAug 4, 2010 ... Meantime GSK has also signed a deal with muscle drug specialist Five Prime Therapeutics that could be worth around $140 million to the San ...

Participants in a University of Florida study use ankle weights to increase strength and balance. Researchers say muscle deterioration is a major reason some of the elderly lose mobility and cannot live independently.

NY Times By ANDREW POLLACK

Published: August 30, 2010

Bears emerge from months of hibernation with their muscles largely intact. Not so for people, who, if bedridden that long, would lose so much muscle they would have trouble standing.

Why muscles wither with age is captivating a growing number of scientists, drug and food companies, let alone aging baby boomers who, despite having spent years sweating in the gym, are confronting the body?s natural loss of muscle tone over time.

Comparisons between age groups underline the muscle disparity: An 80-year-old might have 30 percent less muscle mass than a 20-year-old. And strength declines even more than mass. Weight-lifting records for 60-year-old men are 30 percent lower than for 30-year-olds; for women the drop-off is 50 percent.

With interest high among the aging, the market potential for maintaining and rebuilding muscle mass seems boundless. Drug companies already are trying to develop drugs that can build muscles or forestall their weakening without the notoriety of anabolic steroids. Food giants like Nestlé and Danone are exploring nutritional products with the same objective.

In addition, geriatric specialists, in particular, are now trying to establish the age-related loss of muscles as a medical condition under the name sarcopenia, from the Greek for loss of flesh. Simply put, sarcopenia is to muscle what osteoporosis is to bone.

“In the future, sarcopenia will be known as much as osteoporosis is now,” said Dr. Bruno Vellas, president of the International Association of Gerontology and Geriatrics.

Researchers involved in the effort say doctors and patients need to be more aware that muscle deterioration is a major reason the elderly lose mobility and cannot live independently.

“A doctor sees old people who are shrinking and getting weak, but there is no medical terminology that?s been created and made uniform to allow the doctor to make a diagnosis, look at possible causes, and make a treatment plan,” said Dr. Stephanie A. Studenski, a professor of medicine at the University of Pittsburgh.

Of course, commercial interests are at play as well. “If you are trying to sell drugs, you want to have a very clear criterion for diagnosing the problem and for endpoints to treat it,” said Dr. Thomas Lang of the University of California, San Francisco, who is working on techniques for diagnosing sarcopenia.

A task force of academic and industry scientists met in Rome last November and in Albuquerque last month and has submitted a proposed definition of sarcopenia for publication in a medical journal. The meeting received financial support from several drug companies and food companies.

Underscoring the focus on sarcopenia, four European medical societies proposed a somewhat different definition, and Dr. Studenski is developing yet another.

Whatever the definition, experts say, sarcopenia affects about 10 percent of those over 60, with higher rates as age advances. One study estimated that disability caused by sarcopenia accounted for $18.5 billion in direct medical costs in 2000, equivalent to 1.5 percent of the nation?s health care spending that year.

Causes of the loss of muscle mass or strength might include hormonal changes, sedentary lifestyles, oxidative damage, infiltration of fat into muscles, inflammation and resistance to insulin. Some problems stem from the brain and nervous system, which activate the muscles.

Experts say the best approach to restoring or maintaining muscle mass and strength is exercise, particularly resistance training.

The National Institute on Aging is now sponsoring a controlled trial to test whether exercise can prevent disability in largely sedentary people, age 70 to 89. There is also some early evidence that nutrition, like vitamin D or high levels of protein, might help. “At this point, what we can say is that older people are at risk for eating too little protein for adequate muscle preservation,” said Dr. Elena Volpi of the University of Texas Medical Branch in Galveston.

Pharmaceutical companies are paying more attention to muscles, a part of the body they once largely ignored. A year ago, for instance, GlaxoSmithKline hired William Evans, a leading academic expert on sarcopenia, to run a new muscle research unit.

But with sarcopenia still not established as a treatable condition, “there is no real defined regulatory path as to how one would get approved in this area,” said R. Alan Ezekowitz, a research executive at Merck.

So for now, many companies are focusing on better defined illnesses like muscular dystrophy and cachexia, the rapid muscle wasting that can accompany cancer or other diseases.

One problem is that academic researchers and drug companies initially viewed sarcopenia as primarily a loss of muscle mass, a direct analogy to bone density in osteoporosis. Muscle mass can be measured by the same scans used for bone density.

But some studies have shown that strength, such as gripping force, or muscle function, as measured, say, by walking speed, are more important than mass in predicting problems seniors might have.

“There?s a lot more to the story than simply having a lot of muscle tissue,” said Brian C. Clark, an expert at Ohio University. “Most of the drug stuff has been targeting muscle mass.”

So the definition is shifting to include muscle strength and function. The academic-industry task force recommends testing whether a person can walk four meters, or about 13 feet, in four seconds.

That can be tested by any doctor, without the special equipment needed to measure muscle mass or strength, said Roger A. Fielding of Tufts University, a leader of the task force.

Experts say that to win approval from regulators and reimbursement from insurers, a drug must do more than merely improve mass or strength. It must, for example, improve walking ability or prevent people from falling.

Or perhaps it could restore mobility faster after a person is bedridden. Older people can lose so much muscle during a prolonged hospital stay that they have to move to a nursing home.

Demonstrating such benefits and cost savings would help counter criticism that doctors and drug companies are trying to turn a natural consequence of aging into a disease.

“If you can get out of a nursing home in three weeks instead of three months, wouldn?t we say it is a useful thing?” said Dr. Studenski, who consults for drug companies.

Efforts to develop muscle drugs are still in early stages, and there have been setbacks.

But for inspiration, researchers can look to the bears, though scientists have no definitive answer to the animals? youthful secret.

Moreover, a study that has tracked 3,000 people for 50 years found that about 20 of them, now in their 80s, have not lost muscle mass.

“Maintaining the muscle is possible,” said Dr. Luigi Ferrucci of the National Institute on Aging, who directs the study, called the Baltimore Longitudinal Study of Aging. “We just don?t know the right formula yet.”

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Drug Makers on the Trail of an Alternative to Steroids, to build muscle

NY TimesBy ANDREW POLLACK

Published: August 30, 2010

The hunt is on for alternatives to anabolic steroids, which build muscle by mimicking the effects of the hormone testosterone but whose reputation has been tarnished by athletic doping scandals and side effects like liver damage.

A federally financed study showed that testosterone increased strength in the elderly. But the study was ended abruptly last year because those getting the hormone suffered far more cardiac problems than those getting a placebo.

Human growth hormone is also controversial because of side effects and questions about whether it can increase strength.

Companies like GTx and Ligand Pharmaceuticals are trying to develop drugs that possess the muscle-building ability of testosterone without its side effects, like the development of facial hair and other masculine features in women. These drugs are called selective androgen receptor modulators, or Sarms.

Pfizer, Amgen and Acceleron Pharma are separately pursuing drugs that block myostatin, a protein made by the body that acts as a brake on muscle formation.

Belgian Blue cattle, which do not make myostatin, have huge, rippling muscles and yet are otherwise apparently healthy. Several years ago, scientists reported that a German boy who lacked myostatin because of genetic mutations had abnormal strength and muscle mass.

Cytokinetics is testing a drug for Lou Gehrig?s disease that improves the ability of muscles to contract by, in effect, making them more sensitive to nerve signals.

Most of the drugs have been tested in only early-stage clinical trials so far, and there have been many setbacks.

Wyeth, now part of Pfizer, dropped a myostatin inhibitor that did not work well in a muscular dystrophy clinical trial. Amgen last month called off a trial to test its myostatin blocker for age-related muscle decline. Merck withdrew from a partnership to develop GTx?s drug.

These two phase 2 studies enrolled antiretroviral-naive people with a viral load at or above 5000 copies, a CD4 count above 50, and no resistance to nucleosides, nonnucleosides, or protease inhibitors. Both trials were double-blind and active controlled, the first comparing QUAD with Atripla, the second comparing atazanavir/cobicistat with atazanavir/ritonavir in people also taking tenofovir/emtricitabine.

Researchers randomized 48 people to QUAD, 23 to Atripla, 56 to atazanavir/cobicistat, and 29 to atazanavir/ritonavir. Ages averaged about 35, and about 90% of study participants were men. Median pretreatment viral loads were 4.6 or 4.7 log in all four treatment groups (about 40,000 to 50,000 copies). Median pretreatment CD4 counts were 354 in the QUAD group, 436 in the Atripla group, 341 in the atazanavir/cobicistat group, and 367 in the atazanavir/ritonavir group. AIDS rates were 16% or lower in all treatment arms.

Six people in the atazanavir/cobicistat group never received an antiretroviral dose and were not included in the analysis. Three people (13%) discontinued QUAD, none because of adverse events; 3 people quit Atripla, 1 because of an adverse event (suicidal ideation); 5 people (10%) quit the cobicistat group, 2 because of side events (vomiting and rash); and 3 people (10%) quit the ritonavir group, 1 because of an adverse effect (ocular icterus). Rates of drug-related grade 1 to 4 adverse events were 46% with QUAD, 57% with Atripla, 36% with atazanavir/cobicistat, and 48% with atazanavir/ritonavir. Rates of grade 3 or 4 adverse events were 4% with QUAD, 9% with Atripla, 4% with cobicistat, and 0% with ritonavir.

In a missing-data-equal-failure analysis, proportions with a week-48 viral load below 50 copies were 90% with QUAD versus 83% with Atripla, and 82% with atazanavir/cobicistat versus and 86% with atazanavir/ritonavir. The first result meant QUAD was noninferior to Atripla in 48-week efficacy. In a missing-data-excluded analysis, 48-week sub-50-copy rates were 96% with QUAD versus 95% with Atripla, and 91% with cobicistat versus 96% with ritonavir. CD4 count gains through 48 weeks averaged 240 with QUAD, 162 with Atripla, 230 with atazanavir/cobicistat, and 206 with atazanavir/ritonavir.

Mean percent changes in glomerular filtration rate estimated by the Cockroft-Gault method (eGFR) were -14% with QUAD versus -4% with Atripla and -12% with atazanavir/cobicistat versus -11% with atazanavir/ritonavir. Changes in eGFR appeared early in the trial and remained stable through 48 weeks. Concerns over renal toxicity with cobicistat arose at week 24 in this trial, when eGFR and serum creatinine were worse with QUAD than with Atripla [2]. In the cobicistat-ritonavir comparison, rates of jaundice were low (3% to 4%) and similar in the two treatment arms. Rates of ocular icterus were higher but equivalent in the two arms--12% in the cobicistat group and 14% in the ritonavir group. (from Jules: Gilead has stated that this change in eGFR is not a real toxicity but is difficult to distinguish from one; Gilead?s press release discusses it more as well as the reported study at CROI).

QUAD Four-in-One Pill as Strong as Atripla, But a Kidney Concern ...17th CROI Conference on Retroviruses and Opportunistic Infections San Francisco CA ... Age averaged 36 in the Quad group and 35 in the Atripla group. ...

Clinical Pharmacology at CROI 2010: Advances in Antiretroviral ...Recall that GS-9350 was first reported at the 2009 CROI (Abstract #40) as an agent that ... One study compared the once daily "QUAD" tablet of EVG, GS-9350, ...

Long-term Effects of a Lifestyle Intervention on Weight and Cardiovascular Risk Factors in Individuals With Type 2 Diabetes Mellitus (this can work in HIV for anyone) - pdf attached

Four-Year Results of the Look AHEAD Trial

The Look AHEAD Research Group

Arch Intern Med. Sept 17 2010

"The Look AHEAD trial is, to our knowledge, the first study to examine the effects of an intensive lifestyle intervention through 4 years of follow-up in a large cohort of overweight and obese individuals with type 2 DM.....The average age was 58.7 (6. years, the average BMI was 36.0 (5.9), and the average duration of type 2 DM was 6.8 (6.5) years......The ILI10 included diet modification and physical activity and was designed to induce at least a 7% weight loss at year 1 and to maintain this weight loss in subsequent years....exercise goal was at least 175 minutes of physical activity per week, using activities similar in intensity to brisk walking. Behavioral strategies, including self-monitoring, goal setting, and problem solving, were stressed.....participants in the ILI group experienced greater improvements in weight, fitness, glycemic control, blood pressure, and levels of high-density lipoprotein cholesterol (HDL-C) and triglycerides than those in the DSE group (Table 1). The DSE group experienced greater overall reductions in low-density lipoprotein cholesterol (LDL-C) levels.....Fitness increased by 20.4% in ILI participants and by 5.0% in DSE participants (P < .001) between baseline and year 1. At year 4, the fitness level of ILI participants was still 5.1% over baseline, whereas DSE participants were 1.1% below baseline (P < .001)......A significantly greater proportion of ILI participants met the ADA goal for HbA1c level at each year and for blood pressure at years 1, 2, and 3."

ABSTRACT

Background - Lifestyle interventions produce short-term improvements in glycemia and cardiovascular disease (CVD) risk factors in individuals with type 2 diabetes mellitus, but no long-term data are available. We examined the effects of lifestyle intervention on changes in weight, fitness, and CVD risk factors during a 4-year study.

Methods - The Look AHEAD (Action for Health in Diabetes) trial is a multicenter randomized clinical trial comparing the effects of an intensive lifestyle intervention (ILI) and diabetes support and education (DSE; the control group) on the incidence of major CVD events in 5145 overweight or obese individuals (59.5% female; mean age, 58.7 years) with type 2 diabetes mellitus. More than 93% of participants provided outcomes data at each annual assessment.

Background - Long-term evidence from randomized trials of the effectiveness of exercise in preventing disability and fall-related fractures in elderly people has been lacking.

Methods - We performed extended follow-up of 160 women (aged 70-73 years at baseline) with osteopenia in a population-based, randomized, controlled exercise trial. The trial was conducted from April 1 through April 30, 2001. Follow-up was conducted from May 1, 2001, through December 31, 2005. Mean total time in observation was 7.1 years. Primary outcome measures were femoral neck bone mineral density, postural sway, and leg strength. Secondary outcome measures were hospital-treated fractures and functional ability measures. Outcomes were measured annually using masked assessors.

Research on killer HIV antibodies provides promising new ideas for vaccine designNew discoveries about the immune defenses of rare HIV patients who produce antibodies that prevent infection suggest a novel direction for designing new vaccines. Researchers at Rockefeller University and colleagues have now made two fundamental discoveries about the so called broadly neutralizing anti-HIV antibodies, which effectively keep the virus at bay. By detailing the molecular workings of a proven immune response, the researchers hope their work will ultimately enable them to similarly arm those who are not equipped with this exceptional immunological firepower. The findings are reported in the Sept. 30 issue of the journal Nature.

"Nobody yet can make a vaccine that elicits these broadly neutralizing antibodies, but here are patients who can do it, so let's understand how," says Michel C. Nussenzweig, Sherman Fairchild Professor and head of the Laboratory of Molecular Immunology. "That's the theme in this work. The reason the research community is not making this vaccine is not that we're not good engineers. We are. The reason is that we don't understand how these patients produce these antibodies, and that's what we're figuring out. If we know how they're doing it, we might learn how to reproduce it."

HIV strains mutate rapidly, making them notoriously evasive targets for the immune system. In particular, the HIV envelope spike, called gp160, is the site of a host of mutations that obstruct the few elements that all of the virus strains share. Prior research has shown that only four super antibodies block the activity of that protein in a broad range of HIV strains, neutralizing the virus. But all attempts to coax the human body into producing those four have failed.

Last year, in experiments reported in Nature, the Nussenzweig lab showed that a diverse group of broadly neutralizing antibodies cloned from 433 B cells of six slow progressing HIV patients were as capable of knocking down a broad range of HIV strains as any one of the super antibodies. The ability to isolate and clone antibodies from B cells was first worked out by the lab in a pioneering 2003 paper in Science. Now, having applied that method to the B cells in HIV patients with high titers of broadly neutralizing antibodies, the new research explores in more detail what their antibodies target and how they attack.

Novel discoveries offer new ways to design HIV vaccineThursday, September 30, 2010 4:25:25 AM by ANI ( Leave a comment )London, Sep 30 (ANI): Paving a new way for designing AIDS vaccines, researchers have made novel discoveries about the immune defenses of rare HIV patients who produce antibodies that prevent infection.

Researchers at Rockefeller University and colleagues have now made two fundamental discoveries about the so-called broadly neutralizing anti-HIV antibodies, which effectively keep the virus at bay.

By detailing the molecular workings of a proven immune response, the researchers hope their work will ultimately enable them to similarly arm those who are not equipped with this exceptional immunological firepower.

“Nobody yet can make a vaccine that elicits these broadly neutralizing antibodies, but here are patients who can do it, so let’s understand how,” Nature quoted Michel C. Nussenzweig, Sherman Fairchild Professor and head of the Laboratory of Molecular Immunology, as saying.

“That’s the theme in this work. The reason the research community is not making this vaccine is not that we’re not good engineers. We are. The reason is that we don’t understand how these patients produce these antibodies, and that’s what we’re figuring out. If we know how they’re doing it, we might learn how to reproduce it,” he added.

HIV strains mutate rapidly, making them notoriously evasive targets for the immune system.

In particular, the HIV envelope spike, called gp160, is the site of a host of mutations that obstruct the few elements that all of the virus strains share.

Prior research has shown that only four super antibodies block the activity of that protein in a broad range of HIV strains, neutralizing the virus. But all attempts to coax the human body into producing those four have failed.

Last year, the Nussenzweig lab showed that a diverse group of broadly neutralizing antibodies cloned from 433 B cells of six slow progressing HIV patients were as capable of knocking down a broad range of HIV strains as any one of the super antibodies.

In the new study, the researchers found that most antibodies are traditionally thought to bind to their target, or antigen, in a bivalent fashion, meaning they get a firm grip by taking hold of two specific handles.

But HIV virions do not allow for that possibility because the gp160 spikes are too far apart.

Therefore, antibodies to the virus are handicapped because they can only use one of their two high affinity arms to recognize the viral spike.

The researchers found that on average 75 percent of the anti-gp160-HIV antibodies in their large collection were selected by the immune system for polyreactivity, a property that allowed the second “free” arm of the antibody to enhance overall affinity by binding to the virion “non-specifically.”

Generally, the immune system weeds out polyreactive antibodies, even though they are naturally produced in significant quantities, because polyreactive antibodies could in theory attack the body itself.

But the experiments suggest that these “sticky” antibodies may be an opportunistic adaptation to difficult cases such as HIV, in which homotypic bivalent bonding may not be an option.

Researchers believe that vaccine designed to elicit antibodies that mimic these properties could be a promising strategy to beat the deadly virus.

The study has been published in the latest issue of the journal Nature. (ANI)

The Single-Tablet Regimen Elvitegravir/Cobicistat/Emtricitabine/Tenofovir Disoproxil Fumarate (EVG/COBI/FTC/TDF; "QUAD") Maintains a High Rate of Virologic Suppression, and Cobicistat (COBI) is an Effective Pharmacoenhancer Through 48 Weeks

PHILADELPHIA AIDS researchers announced Thursday that they had finally cracked a long-standing puzzle: Why a few people can get infected with the AIDS virus and remain healthy without treatment?

It was the culmination of a 16-year effort that started with one HIV-positive minister coming into the office of AIDS researcher Bruce Walker and asking to become a human guinea pig.

Episcopal minister Robert Massie was expected to have died from his infection years earlier, and yet felt inexplicably well. He thought if doctors studied him, they might find a way to help others with HIV to stay healthy too.

Eventually, thousands of HIV-positive volunteers joined the effort, helping scientists to pinpoint a set of genetic differences that allow about 1 in 300 infected people to keep the virus in check.

Thanks to their genetics, these "controllers" have a slightly different immune response - a better ability to signal danger so that "killer" T-cells can keep the AIDS virus from replicating and destroying immune cells.

The researchers hope their findings, released Thursday afternoon in the online version of the journal Science, could help inform the quest for new AIDS therapies. About 33 million people worldwide are infected with the virus.

Luis Montaner, an immunology professor at Philadelphia's Wistar Institute, called the work "a very significant and Herculean" achievement. Montaner is engaged in a related project, trying to prompt patients' immune systems to control the virus even when they're not gifted with controller genes.

Massie said he discovered he was HIV positive in 1984, the same year he was married. He was 26 years old. Since childhood, he had been treated for the genetic blood disorder hemophilia, which required him to get frequent blood transfusions. His HIV infection was traced back to a transfusion in 1978, which meant he had been living with the virus for more than five years.

Doctors at the time told him five years was about as long as anyone had carried the virus before getting deathly ill with AIDS. After he had had HIV for 10 years, that became the new outer limit.

"I always had a sense I was kind of staring right off the cliff," Massie said.

By 1994, he knew there was something unusual about him, since nobody was supposed to remain symptom-free for 16 years. A former college roommate who had become a doctor agreed, and after some asking around, Massie connected with Walker at Massachusetts General Hospital.

"He was eager to start doing research on me and asked me a lot of questions," Massie said. "He was very pleased that I lived just a couple of miles from Mass General so he could draw my blood on a regular basis."

Walker, who is now a professor of medicine at Harvard Medical School and director of the Partners AIDS Research Center at Massachusetts General, said he had gotten interested in such cases a few years earlier, after another researcher alerted him to something strange she had seen among a cohort of gay men who had been tested for hepatitis B. Many had also tested positive for HIV, though a few remained symptom-free years after infection.

Walker said at first he assumed Massie had gotten a false positive test result. But retesting proved he was indeed HIV positive. At a conference, Walker quizzed other doctors and found that many of them had seen a small number of similar cases.

Eventually, he said, he found that about 1 in 300 people were "controllers" - able to carry untreated infection without illness.

Those people showed very little of the virus in their blood - in many cases it was undetectable, though they tested positive for HIV antibodies, proving they'd been exposed.

Meanwhile, Walker kept studying Massie, often going to the minister's house to take blood and chat over coffee. "We became really good friends," Massie said. Massie kept asking Walker whether they had any idea what was protecting him from AIDS. For years, the answer was no.

Gradually, Walker's project grew to encompass researchers all over the world and more than 3,500 HIV-positive volunteers, about 1,500 of whom were probable "controllers."

"The patients were absolutely spectacular in terms of their contributions to this," said Walker. "They enthusiastically lined up for this study when they heard about it - some of them flew into Boston to have their blood drawn."

It was a combination of their large volunteer pool and advancing genetics technology that finally allowed them to crack the mystery, said Walker. What they wanted was something these HIV controllers shared that made them different. The human genetic code is 3 billion letters long, Walker said, making it hard to know where to look.

Luckily, by the early 21st century, such studies could take advantage of a group of 3 million genetic signposts, called SNPs, where the human genetic code tends to vary. These can help isolate regions on a chromosome where key differences are likely to be found between one group and another.

Using the SNPs led them to a group of genes on chromosome 6 that code for a component of the immune system called HLA - human leukocyte antigens - which distinguish the body's own cells from foreign invaders.

A further breakthrough pointed to a specific set of these HLA genes where subtle genetic differences changed the way the immune system worked.

Walker describes the HLA proteins as factory workers. When a virus invades a cell, these HLA proteins can grab a piece of the viral protein and "hang it out the window." That, he said, signals that something terrible is going on in that cell. "Then, the killer T cells come and blow up the factory."

In most people, that system doesn't work well against HIV, since the virus attacks key cells in the immune system faster than they can stop it. But in those fortunate few, the HLA proteins do a better job of displaying the viral proteins, which signals infection and prompts the killer T-cells to come in and destroy the infected cell. These so-called controllers showed up among different ethnicities.

Had HIV mutated into a form that was spread by casual contact, these people might make up the majority of the remaining human race. "That's why variability in the human genome is so important," Walker said. The human immune system is a balancing act - and at some being too sensitive would lead to false alarms.

Ian Frank, an AIDS researcher at the University of Pennsylvania, said the many volunteers were crucial to this advance. "That really gives them the power to see things you could never see if you were analyzing five or 10 of these patients," he said, "which is the way most of this research goes."

Massie, who is now 54, used the extra time afforded by his "controller" status to write an award-winning book on apartheid, earn a doctorate in business at Harvard, found a nonprofit that helps companies deal with climate, energy and human rights issues, and run for lieutenant governor of Massachusetts in 1994. He won the primary, he said, making him the first openly HIV-positive candidate for a major office.

Eventually, however, he began getting sick from a latent hepatitis C infection, also picked up from a blood transfusion. His immune system couldn't fight that so he got a liver transplant last year.

He's now feeling well enough to start up his career again. And thanks to the liver transplant, he no longer has the uncontrolled bleeding of hemophilia.

Combination antiretroviral therapy (cART) has led to a major reduction in HIV-related mortality and morbidity; however, HIV can still not be cured. Achieving either a functional cure (long-term control of HIV in the absence of cART) or a sterilizing cure (elimination of all HIV-infected cells) remains a major challenge. The most significant barrier to cure is the establishment of a latent or ‘silent’ infection in resting CD4+ T cells. Several randomized clinical trials have demonstrated that treatment intensification with additional antiretrovirals has little impact on latent reservoirs. Some potential other approaches that may reduce the latent reservoir include very early initiation of cART and the use of agents that could reverse latent infection. Drugs such as histone deacetylase inhibitors, currently used and licensed for the treatment of some cancers; methylation inhibitors; cytokines such as IL-7 or activators of nuclear factor kappa B (NF-κB) such as prostratin, show promising activity in reversing latency in vitro when used either alone or in combination. Alternate strategies include using gene therapy to modify expression of CCR5 and therefore make cells resistant to HIV. This review will primarily focus on the advantages and disadvantages of methods currently being used to quantify persistent virus ex vivo in patients receiving cART and strategies aimed at cure that are being tested in vitro or in early clinical development. In addition, we discuss key issues that need to be addressed to successfully move laboratory research to clinical trials aimed at curing HIV.

Introduction

Despite the significant reduction in morbidity and mortality following combination antiretroviral therapy (cART), cART cannot eradicate HIV. Recently, there has been a renewed scientific interest in developing new strategies to eventually find a cure for HIV. There have been several significant advances in our understanding of the major barriers to curing HIV. These barriers include long-lived latently infected cells and residual viral replication, at least in some patients. In addition, anatomical reservoirs including the gastrointestinal tract, lymphoid tissue and the central nervous system (CNS) may harbour unique long-lived infected cells and penetration of cART may be limited at these sites. The complex mechanisms of how latency is established and maintained in different T-cell subsets and the major cellular reservoirs that persist in patients on cART have recently been extensively reviewed elsewhere [1–5]. In this review, we will therefore focus on the key scientific and clinical variables that we need to understand in order to significantly expand the breadth and scope of clinical trials aimed at finding a cure for HIV. In addition we will focus on strategies for cure that are currently being or soon to be tested in clinical trials.

Why do we need a cure for HIV?

Even with the major successes of cART, full life expectancy for patients living with HIV has not been restored. Although some cohort studies have shown near normal life expectancy for a subset of patients [6], other studies have shown that life expectancy remains shortened [7,8]. In a prospective study of 3990 HIV-infected individuals in Denmark, the chance of a person with HIV reaching the age of 70 was 50% that of uninfected population controls [9]. The incidence of significant morbidity also remains elevated despite successful cART (reviewed in [10]), due to complex interactions between drug toxicity [11], persistent inflammation [12], and risk behaviours [13]. Finally, despite the clear need for universal access to cART, the lack of financial resources to support lifelong treatment, for everyone in need of treatment, is still a major challenge [14,15].

Functional or sterilizing cure?

There are two potential strategies for cure. The first is what could be considered an ‘infectious diseases model’ of cure which would require the elimination of all HIV-infected cells in all compartments and sanctuaries and for patients to have a plasma HIV RNA count of less than 1 copy/ml. This is now commonly referred to as a sterilizing cure. The alternative approach would be to aim for remission or what could be considered a ‘cancer model’ of cure, in which an individual would have long-term health in the absence of treatment, with low-level viraemia at less than 50 copies/ml. This is commonly referred to as a functional cure.

The recent case report of a German patient with acute myeloid leukaemia, who received a bone marrow transplant from a donor who carried a 32-base pair deletion in the CCR5 gene, is the only current example of a sterilizing cure [16]. Following transplantation, the patient stopped cART and HIV RNA remained at below 1 copy/ml. In more detailed studies, including multiple biopsies of his gastrointestinal tract, analysis of his cerebrospinal fluid (CSF) and bone marrow and even a brain biopsy, neither HIV DNA or HIV RNA was detected [16,17]. The patient has now been off cART for over 45 months and HIV is still not detected. Reconstitution of circulating and mucosal CD4+ T cells that did not express CCR5 was observed [17]. CCR5+ macrophages were detected early post transplantation in the gastrointestinal tract but at later time points, all mucosal macrophages expressed the mutant CCR5 [17]. In addition, the patient's peripheral blood mononuclear cells (PBMCs) were permissive to CXCR4 using laboratory isolates ex vivo, demonstrating that the patients CD4+ T cells were not resistant to HIV. Potential factors leading to the elimination of long-lived reservoirs in this patient could have included the specific chemotherapy administered, total body irradiation or low-grade graft-versus-host disease in addition to eliminating the capacity for any residual replication by removing target cells that express CCR5. Whereas a strategy of using bone marrow transplantation with a CCR5 mutant donor is not a realistic cure for HIV given the toxicity and complexity of the treatment, we need to continue to comprehensively study this patient to fully understand how and why HIV was eliminated.

Functional cure: elite controllers

Elite controllers represent a unique group of patients who are able to achieve a consistent and long-term control of viral replication with HIV RNA of less than 50 copies/ml in the absence of cART. In addition, the reservoir is significantly smaller in elite controllers with low concentration of HIV DNA in different subsets of circulating CD4+ T cells in blood [18,19] as well as in rectal tissue [20,21].

There have been multiple studies examining the role of genetics, the virus and the immune response in elite controllers [20,22–25]. One of the consistent results from this work is the clear association with HLA class one genes [26,27]. Recent work has also demonstrated the importance of an effective cytolytic CD8+ T-cell response in blood which has been associated with enhanced activity of the T-box transcription factor t-bet [28,29] and increased production of IL-21 [30]. Strong HIV-specific CD4+ and CD8+ T-cell responses were also identified in mucosal tissue from elite controllers [31,32]. The innate immune system may also be important with enhanced activity of myeloid dendritic cells [33]. These data provide supportive evidence that inducing an effective immune response, perhaps via vaccination, may be one strategy to achieve a functional cure.

As some elite controllers do not bear the protective alleles HLA B27 or HLA B57, mechanisms other than enhanced T-cell immunity have also been explored. Several investigators have demonstrated lower replicative capacity of the virus isolated from elite controllers [34–36], and very low level of viral replication soon after infection [37]. There is no evidence currently that activated CD4+ T cells from these patients are resistant to HIV [38].

Despite apparent ‘functional cure’ in elite controllers, it is important to remember that low-level viraemia and infected resting CD4+ T-cells are detected [23,24]. Compared with patients receiving cART, PBMC from elite controllers have similar levels of total DNA, but significantly lower integrated DNA and higher 2-long terminal repeat (2-LTR) levels [39]. Immune activation is higher in elite controllers compared with healthy controls [22]. In contrast to patients on cART with HIV RNA below 50 copies/ml, there is evolution in HIV RNA sequences in elite controllers [40], and in approximately 7% of elite controllers, CD4+ T cells decline over time [24]. Because of the low total number of infected cells and robust HIV-specific immune responses, elite controllers could potentially be the best candidates to test strategies aimed at achieving a sterilizing cure.

Measuring latently infected cells and the ‘reservoir’ in vivo

The major reason why HIV cannot be cured is the persistence of HIV in a latent form in different cellular reservoirs. In vivo, HIV latency occurs in resting CD4+ T cells either as preintegration or postintegration latency. Preintegration latency refers to unintegrated HIV DNA that is unstable and will either degrade or will integrate into the host cell genome, usually following cell activation [41]. Postintegration latency refers to the presence of integrated HIV DNA in cells that are not actively producing viral particles [42].

The major reservoir of cells that harbour postintegration latency in vivo are resting central memory (CD45RA-CCR7+CD27+) and transitional memory (CD45RA-CCR7-CD27+) CD4+ T cells [43,44]. Latent infection can also be established in other long-lived cells including naïve T cells [45,46], bone marrow progenitor cells [47], thymocytes [48], and astrocytes [49,50]. Other cells such as monocyte/macrophages can support long-lived low-level productive infection [51].

Together, these persistent infected cells constitute the ‘latent reservoir’. Latently infected cells can be detected in both blood and tissue, including the gastrointestinal tract [52], genital tract [53] and the central nervous system [50,54]. When activated, latently infected T cells can either release viral particles or became productively infected T cells. In the presence of treatment, further rounds of infection do not occur and there is no viral rebound but when treatment is stopped, viral rebound will occur. There are multiple methods currently used to quantify persistent HIV-infected cells in patients on cART (summarized in Fig. 1 and Table 1) [39,42,53,55–73].